Single-particle reconstruction (SPR) from electron microscopy (EM) images is widely used in structural biology, but it lacks direct information on protein identity. To address this limitation, we developed a computational and analytical framework that reconstructs and coaligns multiple proteins from 2D super-resolution fluorescence images. To demonstrate our method, we generated multicolor 3D reconstructions of several proteins within the human centriole, which revealed their relative locations, dimensions and orientations.
Sieben, C., Banterle, N., Douglass, K. M., Gönczy, P., & Manley, S. (2018). Multicolor single-particle reconstruction of protein complexes. Nature Methods, 15(10), 777–780. https://doi.org/10.1038/s41592-018-0140-x